二氧化硅诱导气道上皮炎症中Aurora A激酶介导的信号传导机制研究

Silicosis is an incurable and disabling occupational disease. Our previous work found that Aurora A kinase was highly expressed and greatly correlated with inflammatory factors in silica-induced airway epithelial cells, but the mechanism remains unclear. Since both the p53 and NF-κB pathways promote the expression of inflammatory cytokines and the Aurora A kinase activity increases, it is speculated that Aurora A kinase may affect inflammation by regulating the p53 and NF-κB pathways. This project intends to apply dimethyl labeling-based and multiple reaction monitoring-based quantitative proteomic technology to detect the target proteins and their phosphorylation levels associated with Aurora A kinase, to describe the protein-protein interaction sites, to quantitatively analyze the Aurora A-mediated protein phosphorylation and to systematically elucidate the activation and signal transduction regulation of Aurora A kinase via the p53 and NF-κB pathways. We further investigate the specificity and molecular mechanism of Aurora A kinase in the inflammatory response in silicosis mice. This study would clarify the molecular mechanism of Aurora A kinase in silica-induced airway epithelial inflammation, and provide new ideas for the clinical diagnosis and treatment of silicosis.

矽肺是一种难治愈的致残性职业性疾病。我们前期工作发现由二氧化硅诱导气道上皮细胞产生炎症过程中Aurora A激酶高表达并与炎症因子显著相关，但其机制尚未明了。基于p53和NF-κB通路促进炎症因子表达同时Aurora A激酶活性增加，我们推测Aurora A激酶可能通过调节p53和NF-κB通路影响炎症。本项目拟采用二甲基标记和多重反应监测的定量蛋白质组学技术，检测炎症过程中与Aurora A激酶相关的靶向蛋白质及其磷酸化，明确蛋白质间的相互作用位点，定量分析Aurora A激酶介导的蛋白质磷酸化水平，系统研究p53和NF-κB通路上Aurora A激酶活化和信号传导的调节。与此同时，进一步研究Aurora A激酶参与和主导矽肺病小鼠炎症反应的特异性和分子机理。本研究将阐明Aurora A激酶在二氧化硅诱导气道上皮炎症中的分子机制，为矽肺病的临床诊断和治疗提供新的思路。

本项目针对矽肺病早期二氧化硅会引起上皮细胞产生炎症作为切入点，运用组学技术和分子生物学技术，深入阐述该炎症过程，同时探究抑制剂和蛋白质的相互作用，期望能在临床上为矽肺病的诊断和治疗提供参考和数据支持，从而提高职业病患者的生活质量。本项目在以下三个方面展开并取得如下成果：1）运用代谢组学，蛋白组学及其磷酸化蛋白组学等多组学的技术分析二氧化硅刺激上皮细胞产生炎症的过程，系统分析代谢物，蛋白质及其磷酸化的变化情况，阐述MAPK信号通路的重要作用，并发现上皮细胞的有氧糖酵解的增强；2）Aurora A激酶在该过程高表达，与其相互作用的蛋白p53通过结合IL-8的启动子NC_000004.12调控IL-8的表达从而参与了炎症过程；3）在二甲基标记的基础上又发展了尺寸选择蛋白酶切方法，分析抑制剂结合Aurora A激酶后蛋白的构象变化，从而为药物的设计和开发提供强有力的工具。以上数据可为矽肺病早期的炎症过程提供数据参考并为后续抑制剂的开发提供技术支持。目前本项目已经发表SCI论文4篇（其中1篇刚接收还未在线），其中申请人作为第一作者1篇，第一作者且是通讯作者2篇，其他作者论文1篇，基本完成了项目的预定目标。